Objective To study the effect of inflammatory cytokines on diabetic nephropathy.Methods Blood glucose(BG),blood urea nitrogen(BUN),serum creatinine(Scr) and 24 hour urinary protein(24hUPro) were detected at the end of 4 weeks and 8 weeks study in male SD rats.The rats were divided into diabetes(DM) group and normal control(NC) group.Kidney weight/body weight ratio(KI) of rats were measured and calculated.Kidney pathological changes were observed by light microscope and electron microscope.Expressions of interleukin-18(IL-18),intercellular adhesion molecule-1(ICAM-1)and tumor necrosis factor-alpha(TNF-?) in glomeruli were detected by immunohistochemical staining.Results Compared with homochronous NC group,BG,KI,24hUPro,Scr and expressions of IL-18,ICAM-1,TNF-? were significantly higher in DM group(P

Objective To explore the relationship between the microsatellite markers on chromosome 6 and the phenotypes of atopy. Methods Sixteen microsatellite markers on chromosome 6 were evaluated in 20 affected sib pair and trios families. Linkage disequilibrium analysis was conducted according to asthmatic phenotypes (total IgE, skin prick test, bronchial hyper responsiveness and eosinophil) by ETDT software system. Results Significant P value( P

The serum level of 25-dihydroxyvitamin D3 was determined in patients with type 2 diabetes,diabetic nephropathy,and healthy objects..The results demonstrated that the serum 25-dihydroxyvitamin D3 level in type 2 diabetic patients with nephropathy was lower than that in the healthy objects or type 2 diabetes objects.Blood β2-M,total cholesterol,creatinine,24 h urinary albumin,and course of illness in type 2 diabetic patients with nephropathy have relationship with serum 25-dihydroxyvitamin D3.

BACKGROUND: Islet transplantation is an effective method for the treatment of type 1 diabetes mellitus and parts of type 2 diabetes mellitus. However, its application is hindered by insufficient sources and immunologic rejection. Though transdifferentiation of pancreatic stem cells is at the starting step, it is thought to be the hopeful source for islet cell transplantation.OBJECTIVE: To look for a suitable cells-transplantation source for the treatment of diabetes mellitus. METHODS: The pancreatic ductal epithelial cells were separated from Kunming mice and cultured in DMEM/F12 medium supplemented with keratinocyte growth factor, hepatocyte growth factor and nicotinamide, etc. Samples were taken at different time points for light microscopy and electron microscope. The changes of CK-19 and PDX-1 were detected by immunocytochemistry at 1 and 16 days. The expressions of insulin and glucagon gene were detected by RT-PCR at 1 and 16 days. The physiologic function of these islet-like clusters was determined by dithizone staining and glucose stimulation at 21 days. RESULTS AND CONCLUSION: A large number of epitheliod cells were CK-19 immunoreactive positive and few of them were PDX-1 positive at 1 day after isolation, then CK-19 positive cells proliferated quickly and formed substantial plaques of epithelial cells in cobblestone pattern. At 16 days later, these cells begin to form islet-like clusters gradually, while most of them were PDX-1 immunoreactive positive. The analysis of mRNA by RT-PCR showed very low levels of insulin and glucagon mRNA in the starting materials but increase was found as the process of transdifferentiation. At 21 day differentiated islet-like clusters were stained red by dithizone. In those samples exposed to a stimulatory 15 mmol/L glucose, there was a 1.6-fold increase in insulin compared with to 5.6 mmol/L glucose (P < 0.05). Pancreatic ductal cells of adult Kunming mice could proliferate quickly and have the potency of transdifferentiation into islet-like clusters when cultured in vitro under appropriate conditions.

Streptozotocin-induced diabetic rats were treated with resveratrol for 12 weeks.Compared with group of diabetic rats without treatment,the levels of urine albumin,serum creatinine,and blood urea nitrogen were significantly decreased and pathological changes in kidney were improved in treatment group ( P＜0.05 ).The mRNA expressions of FoxO1 and catalase were higher and FoxO1 phosphorylation level in diabetic rats treated with resveratrol was lower than that in diabetic rats without treatment (P＜0.05),suggesting that resveratrol may protect the kidneys of diabetic rats via regulating expression of FoxO1.

Objective To investigate protective effects of pyrrolidine dithiocarbamate (PDTC) and mycophenolate mofetil (MMF) on kidneys of diabetic rats and the possible mechanism. Methods Thirty-four streptozotocin-induced diabetic rats were randomly divided into 4 groups: diabetic aminals without treatment (D group), diabetic rats treated with PDTC (P group), diabetic rats treated with MMF (M group), diabetic rats treated with combined PDTC and MMF (P+M group), and normal rats were considered as control group (C group). Blood glucose, blood urea nitrogen (BUN), serum creatinine (Scr), and 24 h urinary protein (24 h UP) were detected at the end of treatment lasting for 8 weeks. Kidneys were weighed in order to measure kidney weight/body weight ratio (KI) after rats were killed. Pathological changes in the kidneys were observed by light microscope and electron microscope. Expressions of interleukin-18 (IL-18), intercellular adhesion molecule-1 (ICAM-1) and tumor necrosis factor-α (TNF-α) in renal glomerulus were detected by immunohistochemical staining.Results Blood glucose, BUN, Scr, 24 h UP, KI were significantly higher in D, P, M and P+M groups than those in C group (all P＜0.05). BUN, Scr, 24 h UP, KI were significantly lower in P, M and P+M groups than those in D group (all P＜0.05). 24 h UP was reduced in P+M groups compared with P,M groups (P＜0.05). Pathological changes were attenuated in P,M and P+M groups compared with D group. Expressions of IL-18, ICAM-1, TNF-α in renal glomerulus were much higher in D group than those in C group (all P＜0.05) and were reduced in P, M and P+M groups compared with D group (all P＜0.05). Expressions of IL-18, ICAM-1, TNF-α in P+M group were lower than those in P group (all P＜0.05), while expressions of IL-18, TNF-α were lower than those in M group (both P＜0.05). Conclusion The protective effects on kidneys of diabetic rats produced by combined use of PDTC with MMF were much better than by the application of either PDTC or MMF alone. The mechanism appears to be related with suppressing expressions of inflammatory cytokines.

To study the effects of forkhead transcription factor O1 (FoxO1) on the expression of type Ⅳ collagen and desmin in podocytes of diabetic rats.Streptozotocin-induced diabetic rats were divided into three groups:diabetic rats (DM group),rats transfected with blank lentiviral vectors (diabetes mellitus plus LV-pSC-GFP group,LV-NC group),and rats which were transfected with lentiviral vectors carrying constitutively active FoxO1 (diabetes mellitus plus LV-FoxO1-AAA group,LV-CA group).Rats which received an injection of diluent buffer served as normal control.At 2,4,and 8 weeks after transfection,the levels of urine albumin,blood glucose,blood urea nitrogen,and serum creatinine were measured.Realtime PCR and Western blotting were performed to measure the mRNA and protein levels of FoxO1,COL4A3,COL4A5,and desmin in the renal cortex.Moreover,light microscope and electron microscope were used to observe the structural changes in glomerulus and podocytes.Compared with LV-NC and DM group,in LV-CA group,there was a significant increase in the mRNA and protein levels of FoxO1,and a distinct decrease in the levels of urine albumin,blood urea nitrogen and serum creatinine of rats (except at the twoweek time point) (all P＜0.05),the mRNA and protein levels of COL4A3,COL4A5,and desmin were all decreased (all P＜0.05),and pathological changes in kidney were also improved.Upregulating the expression of FoxO1 by transfecting with constructed lentiviral vectors can definitely improve the abnormal expression of type Ⅳ collagen and desmin in podocytes of diabetic rats.

Objective To observe the clinical effect of compound danshen dropping pills on the early diabetic kidney disease.Methods A total of 850 type 2 diabetes mellitus (T2DM) patients with diabetic nephropathy (DN) at stage Ⅲ who met the criteria for DN from January 2012 to July 2012,aged 60 to 75 years [mean age (66.6±4.8) years],were retrospectively analyzed and randomized into the treatment group (n=601) and the control group (n=249).All patients received rational hypoglycemic,antihypertensive,lipids-adjusting treatment and the treatment group received compound danshen drop pill (15 pills,tid),additionally.Laboratory indexes such as urine albumin (UA) and urine creatinine (UCR) were examined 24 weeks after treatment.Results The levels of UA and UCR,and the UA/UCR ratio (ACR) in the treatment group were decreased after versus before treatment.The decreases of UA and UCR had significant differences between the treatment and control group [(39.1±78.7) mg/L vs.(14.7±77.1) mg/L,(1.1±4.5) mmol/L vs.(-0.3± 3.6) mmol/L,t=4.15 and 4.86,both P＜0.01].Conclusions The comprehensive treatment combined with compound danshen dropping pills for 24 weeks can more effectively reduce UA and ACR,thereby it has effects in treating DN and delaying the progression of DN.

Objective To study the effects of inhibiting forkhead transcription factor O1 (FoxO1)expression by small interference RNA (siRNA) on the glucose utilization of insulin resistant HepG-2 cell line and the mechanism.Methods FoxO1 gene-targeted siRNA vector,which carried a red fluorescence,was constructed and then confirmed by DNA sequencing.HepG-2 cells were induced to a status of insulin resistance by being exposed to 10-6 mol/L insulin for 24 h.The study included 4 groups: HepG-2 cell group cultured with normal medium (group A) ; insulin resistant HepG-2 cell group (group B) ; insulin resistant HepG-2 cell group into which FoxO1 siRNA vector was transfected (group C) ; insulin resistant HepG-2 cell group into which Lipofectamine2000 was added (group D).The transfection efficiency could be estimated by observing the expression of red fluorescence.The expression of FoxO1 mRNA was analyzed by RT-PCR.The expressions of IRS-2 and tyrosine phosphorylation were detected by Western blot and immunoprecipitation.Results FoxO1 gene-targeted siRNA vector was built successfully.The expression of red fluorescence was the strongest at 48 h after transfection.Compared with group A,the glucose consumption and the expression of IRS-2 tyrosine phosphorylation of group B were decreased (P<0.01),and expression of FoxO1 mRNA was increased (P<0.05) in group B.There was no difference in the expressions of IRS-2 protein between A and B groups.Compared with group B,the expression of FoxOl mRNA was decreased (P < 0.01),and the glucose consumption and expression of IRS-2 tyrosine phosphorylation were significantly increased (P<0.05) in group C.There was no difference between group D and group B in glucose consumption, FoxO1 mRNA, IRS-2 protein and IRS-2 tyrosine phosphorylation expressions.Conclusions Inhibiting the expression of FoxO1 gene in insulin resistant HepG-2 cells seems to improve insulin sensitivity by feedback regulating the IRS-2 tyrosine phosphorylation expressions.

Total 306 patients with early stage diabetes were enrolled in the study, after follow-up for 67 ± 8 months, the data of 288 cases were analyzed with survival analysis method. At the end of study 64 out 288 patients had high serum uric acid and the remaining 224 had normal uric acid levels. The prevalence of dominant proteinuria and reduced GFR were significantly higher in patients with high serum uric acid than those with normal serum uric acid (47. 36% vs 30. 29% and 66. 14% vs 37.72%, respectively, both P <0. 05). The results suggest that high level of serum uric acid is a risk factor for the progress of diabetic nephropathy.